We studied the metabolism of polycyclic aromatic hydrocarbons (PAHs) by using white rot fungi previously identified as organisms that metabolize polychlorinated biphenyls. Bran flakes medium, which has been shown to support production of high levels of laccase and manganese peroxidase, was used as the growth medium. Ten fungi grown for 5 days in this medium in the presence of anthracene, pyrene, or phenanthrene, each at a concentration of 5 μg/ml could metabolize these PAHs. We studied the oxidation of 10 PAHs by using laccase purified from Coriolopsis gallica. The reaction mixtures contained 20 μM PAH, 15% acetonitrile in 60 mM phosphate buffer (pH 6), 1 mM 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulfonate) (ABTS), and 5 U of laccase. Laccase exhibited 91% of its maximum activity in the absence of acetonitrile. The following seven PAHs were oxidized by laccase: benzo[a]pyrene, 9-methylanthracene, 2-methylanthracene, anthracene, biphenylene, acenaphthene, and phenanthrene. There was no clear relationship between the ionization potential of the substrate and the first-order rate constant (k) for substrate loss in vitro in the presence of ABTS. The effects of mediating substrates were examined further by using anthracene as the substrate. Hydroxybenzotriazole (HBT) (1 mM) supported approximately one-half the anthracene oxidation rate (k = 2.4 h−1) that ABTS (1 mM) supported (k = 5.2 h−1), but 1 mM HBT plus 1 mM ABTS increased the oxidation rate ninefold compared with the oxidation rate in the presence of ABTS, to 45 h−1. Laccase purified from Pleurotus ostreatus had an activity similar to that ofC. gallica laccase with HBT alone, with ABTS alone, and with 1 mM HBT plus 1 mM ABTS. Mass spectra of products obtained from oxidation of anthracene and acenaphthene revealed that the dione derivatives of these compounds were present.

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White rot fungi from the University of Alberta Mold Herbarium, identified as able to degrade aromatics from a study of PCB metabolism, were examined for production of ligninolytic enzymes. Production of lignin peroxidase, manganese peroxidase, laccase, and veratryl alcohol oxidase were monitored during growth in different media. Good growth but low enzyme production occurred in a glucose - malt extract - yeast extract medium. Media containing 2% cereal bran in 60 mM phosphate buffer supported high levels of laccase production, up to 13 000 U/L in Coriolopsis gallica UAMH 8260 and manganese peroxidase activity up to 1100 U/L in Bjerkandera adusta UAMH 8258. Cereal bran media supported higher laccase production than 2,5-xylidine and higher manganese peroxidase production than a medium containing manganous ion plus veratryl alcohol.Key words: cereal bran, laccase, manganese peroxidase, white rot fungi.

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TITLE OF PROJECT cep+l oc\s o.bour cart'a!J /0 per-eqr{y I The project and the manuscript have been successfully completed and meet the standards of the School ofNursing University. The project demonstrates the application of professional !mow ledge, clinical expertise, and scholarly thinking. An abstract of the project and two copies of the manuscript are attached..